The present invention relates to novel anilide compounds and pharmaceutical compositions comprising them. More precisely, the present invention relates to compounds of a general formula (I): 
wherein Ar represents an optionally-substituted aryl group; R4 and R5 are the same or different, and each represents a hydrogen atom, a lower alkyl group, or a lower alkoxy group; and R4 and R5 may together form a lower alkylene group of which one or more methylene moieties may optionally be substituted by oxygen and/or sulfur atoms;
X represents xe2x80x94NHxe2x80x94, or an oxygen or sulfur atom;
Y represents xe2x80x94NHxe2x80x94, an oxygen or sulfur atom, or a sulfoxide or sulfone group;
Z represents a single bond, or xe2x80x94NR6xe2x80x94;
R6 represents a hydrogen atom or a lower alkylene group; and
n represents an integer of from 0 to 15;
provided that, when X and Y are sulfur atoms, R4 and R5 are hydrogen atoms, Z is a single bond and n is 0, then Ar must not be a phenyl or p-chlorophenyl group, when X and Y are sulfur atoms, R4 and R5 are hydrogen atoms,
Z is a single bond and n is 1, then Ar must not be a phenyl group, and
when X is an oxygen atom, Y is a sulfur atom, R4 and R5 are hydrogen atoms, Z is a single bond and n is 1, then Ar must not be a phenyl group,
their salts and solvates, and pharmaceutical compositions comprising said compounds.
With the recent change in the Japanese eating habits into Western-style ones to take high-calorie and high-cholesterol foods and drinks, which is based on the improvement in the living standard in Japan, and with the recent increase in the aged population of Japan, cases of hyperlipemia and arteriosclerotic disorders resulting from hyperlipemia are greatly increasing with bringing about one social problem in Japan. The conventional chemotherapy for cases of hyperlipemia and arteriosclerosis is essentially to lower their blood-lipid levels that participate in the disorders, but is not targeted to the focuses themselves of arteriosclerosis to cure them.
Acyl coenzyme A cholesterol acyltransferase (ACAT) is an enzyme to catalyze the transfer of cholesterol into cholesterol esters, while playing an important role in the metabolism of cholesterol and the absorption thereof through digestive systems. It is believed that the inhibition of such an ACAT enzyme that may catalyze the esterification of free cholesterol in epithelial cells in small intestines brings about the inhibition of the cholesterol absorption through intestinal tubes, while the inhibition of the formation of cholesterol esters in the liver based on the ACAT inhibition brings about the inhibition of the VLDL (very low-density lipoprotein) secretion into blood, thereby resulting in the decrease in the blood cholesterol. Many known ACAT inhibitors are expected to act on ACAT in small intestines and the liver as anti-hyperlipemic agents thereby to lower blood cholesterol.
For example, as ACAT inhibitors, U.S. Pat. No. 4,716,175 discloses 2,2-dimethyl-N-(2,4,6-trimethoxyphenyl) dodecanamide, and European Patent 372,445 discloses Nxe2x80x2-(2,4-difluorophenyl)-N-[5-(4,5-diphenyl-1H-imidazol-2-ylthio)pentyl]-N-heptylurea. However, any known ACAT inhibitors have heretofore been specifically directed to the decrease in blood cholesterol as anti-hyperlipemic agents, and administered to patients in large amounts in order to express their effects. Therefore, in the clinical examination stage using them, many patients have experienced various side effects such as bleeding from their intestinal tubes, intestinal disorders, diarrhea and liver disorders, which have made it difficult to develop the clinical use of ACAT inhibitors.
WO92/09582 discloses compounds having a certain substituent at the 2-position of the imidazole skeleton) and EP-A 477,778 discloses compounds having certain substituents at the 4- and 5-positions of the imidazole skeleton. For example, disclosed are 5-[2-(2-(4-fluorophenyl)ethyl)-3-(1-methyl-1H-imidazol-2-yl)-2H-benzopyran-6-yl]oxy-2,2-dimethyl-N-(2,6-diisopropylphenyl)pentanamide (see WO92/09582), N-(2,6-diisopropylphenyl)-2-(tetradecylthio)acetamide (see JP 92-500533, WO92/09572), N-butyl-Nxe2x80x2-[2-(3-(5-ethyl-4-phenyl-1-yl)propoxy)-6-methylphenyl]urea (see EP 477,778), and N-[5-(4,5-diphenyl-1H-imidazo-2-ylthio)pentyl]-N-heptyl-2-benzoxazolamine (see WO93/23392); and it is disclosed that these compounds have ACAT inhibiting activities. However, the chemical structures of these compounds are quite different from those of the compounds of the present invention.
3-(benzothiazol-2-ylthio)-N-(phenyl)propanamide, 3-(benzothiazol-2-ylthio)-N-(phenyl)ethanamide and 3-(benzothiazol-2-ylthio)-N-(p-chlorophenyl) ethanamide is reported in J. Chem. Eng. Data, 27, 207 (1982) and 3-(benzoxazol-2-ylthio)xe2x80x94Nxe2x80x94(phenyl)propanamide is reported in Fungitsidy, Ed. Melnikov, N. N. Izd. Fan Uzb. SSR: Tashkent, USSR, 82-88 (1980)).
Naturally, arteriosclerosis is a disorder that is characterized by the increase in the thickness of intimate and the accumulation of lipids in blood vessels. The recent studies on this disorder, arteriosclerosis have clarified that the inhibition of macrophage-derived foam cells that play the central role in the formation of focuses of arteriosclerosis is expected to reduce the focuses themselves of arteriosclerosis. In the focuses of atheromatous arteriosclerosis, seen are macrophage-derived foam cells (which have fatty drops of cholesterol esters therein), and it is said that the formation of such foam cells from macrophages has close relation to the growth of the focuses of arteriosclerosis. In addition, it has been reported that the ACAT activity in the blood vessel walls in the site with arteriosclerotic lesions is increased and that cholesterol esters are accumulated on the blood vessel walls in said site (see Gillies, P. J. et al.; Exp. Mole, Pathol., 44, 329-339 (1986)).
Since the inhibition of the esterification of cholesterol by an ACAT inhibitor produces free cholesterol in cells while the resulting free cholesterol is removed from the cells by a high-density lipoprotein (HDL) and brought to the liver (countertransference of cholesterol via HDL) and metabolized therein, it is expected that such an ACT inhibitor may inhibit the accumulation of cholesterol esters in the site of arteriosclerotic lesions. As a result, it is believed that ACAT inhibitors exhibit direct anti-arteriosclerotic effects. It has been reported that ACAT includes two sub-types, one existing in small intestines and the other existing in blood vessel walls (see Kinnunen, P. M., et al.; Biochem., 27, 7344-7350 (1988) ). Many studies on ACAT inhibitors have heretofore been made for the former sub-type of ACAT existing in small intestines and the liver (see Tomoda, H. et al.; J. Antibiotics, 47, 148-153 (1994)). Having considered that chemicals capable of selectively inhibiting the latter sub-type of ACAT existing in blood vessel walls could be medicines with few side effects for curing arteriosclerosis, as compared with ACAT inhibitors not specific organs, we, the present inventors have searched for inhibitors for ACAT of that type and have studied to synthesize such. ACAT inhibitors.
In order to attain this object, we have made various studies and, as a result, have found that compounds of a general formula (I): 
wherein Ar represents an optionally-substituted aryl group;
R4 and R5 are the same or different, and each represents a hydrogen atom, a lower alkyl group, or a lower alkoxy group; and R4 and R5 may together form a lower alkylene group of which one or more methylene Moieties may optionally be substituted by oxygen and/or sulfur atoms;
X represents xe2x80x94NHxe2x80x94, or an oxygen or sulfur atom;
Y represents xe2x80x94NHxe2x80x94, an oxygen or sulfur atom, or a sulfoxide or sulfone;
Z represents a single bond, or xe2x80x94NH6xe2x80x94;
R4 represents a hydrogen atom or a lower alkylene group; and
n represents an integer of from 0 to 15;
provided that, when X and Y are sulfur atoms, R4 and R5 are hydrogen atoms, Z is a single bond and n is 0, then Ar must not be a phenyl or p-chlorophenyl group,
when X and Y are sulfur atoms, R4 and R5 are hydrogen atoms,
Z is a single bond and n is 1, then Ar must not be a phenyl group, and
when X is an oxygen atom, Y is a sulfur atom, R4 and R5 are hydrogen atom, Z is a single bond and n is 1, then Ar must not be a phenyl group,
and their salts and solvates have excellent ACAT inhibiting activities. On the basis of these findings, we have completed the present invention.
We, the present inventors have found that some of those compounds have organ-selective, ACAT inhibiting activities and intracellular cholesterol transference inhibiting activities, as well as excellent blood cholesterol-reducing activities, and are therefore useful as anti-hyperlipemic agents, while some others have activities to inhibit the formation of foam cells from macrophages and are therefore especially useful as medicines for preventing and curing arteriosclerosis
Accordingly, the present invention provides compounds of the a above-mentioned formula (I), and their salts and solvates.
In addition, the present invention also provides pharmaceutical compositions comprising any of compounds of the above-mentioned formula (I) and their salts and solvates, along with pharmaceutically-acceptable carriers.
The present invention further provides ACAT inhibitors, intracellular cholesterol transference inhibitors, blood cholesterol depressants, and inhibitors for macrophage foam cells which comprise compounds of a general formula (III): 
wherein Ar represents an optionally-substituted aryl group;
R4 and R5 are the same or different, and each represents a hydrogen atom, a lower alkyl group, or a lower alkoxy group; and R4 and R5 may together foam a lower alkylene group of which one or more methylene moieties may optionally be substituted by oxygen and/or sulfur atoms;
X represents xe2x80x94NHxe2x80x94, or an oxygen or sulfur atom;
Y represents xe2x80x94NHxe2x80x94, an oxygen or sulfur atom, or a sulfoxide or sulfone;
Z represents a single bond, or xe2x80x94NR6xe2x80x94;
R6 represents a hydrogen atom or a lower alkylene group; and
n represents an integer of from 0 to 15;
and their pharmaceutically-acceptable salts and solvates, optionally along with pharmaceutically-acceptable carriers. Specifically, the present invention provides medicines for treating, preventing and curing disorders of, for example, hyperlipemia, arteriosclerosis, cervical and cerebral arteriosclerosis, cerebrovascular disorders, ischemic cardiopathy, coronary arteriosclerosis, nephrosclerosis, arteriosclerotic nephrosclerosis, arteriolosclerotic nephrosclerosis, malignant nephrosclerosis, ischemic enterophathy, acute mesenteric vaso-obstruction, chronic intestinal angina, ischemic colitis, aortic aneurysm, and arteriosclerosis obliterans (ASO), which medicines comprise any of compounds of the above-mentioned formula (III) and their pharmaceutically-acceptable salts and solvates, optionally along with pharmaceutically-acceptable carriers.
The present invention further provides a method for treatments using an ACAT inhibitor, an intracellular cholesterol transference inhibitor, a blood cholesterol depressant) or an anti-foaming agent for macrophages, which comprises an effective dosage of any of compounds of a general formula (III) and their salts and solvates.
The present invention provides further also use for the preparation of medicines using an ACAT inhibitor, an intracellular cholesterol transference inhibitor, a blood cholesterol depressant, or an anti-foaming agent for macrophages, which comprises an effective dosage of any of compounds of a general formula (III) and their salts and solvates.
The aryl group for Ar in formula (I) may include a phenyl group, an xcex1-naphthyl group and a xcex2-naphthyl group, and is preferably a phenyl group. The group Ar may be substituted by any substituents not having any negative influence on the ACAT inhibiting activities of compounds (I). Preferred substituents for Ar are, for examples a lower alkyl group, a lower alkoxy group, a halogen atom, a hydroxyl group, a phosphoric acid group, a sulfonamido group, and an optionally-substituted amino group. Especially preferred are a lower alkyl group, a lower alkoxy group and a halogen atom.
More precisely, the present invention provides compounds of a formula (II): 
wherein R1, R2 and R3 are the same or different, and each represents a hydrogen atom, a lower alkyl group, a lower alkoxy group, a halogen atom, a hydroxyl group, a phosphoric acid group, a sulfonamido group, or an optionally-substituted amino group; or any two of R1, R2 and R3 together form an alkylenedioxy group; R4 and R5 are the same or different, and each represents a hydrogen atom, a lower alkyl group, or a lower alkoxy group; and R4 and R5 may together form a lower alkylene group of which one or more methylene moieties may optionally be substituted by oxygen and/or sulfur atoms;
X represents xe2x80x94NHxe2x80x94, or an oxygen or sulfur atom;
Y represents xe2x80x94NHxe2x80x94, an oxygen or sulfur atom, or a sulfoxide or sulfone;
Z represents a single bond, or xe2x80x94NR6xe2x80x94;
R6 represents a hydrogen atom or a lower alkylene group; and
n represents an integer of from 0 to 15;
provided that, when X is an oxygen or sulfur atom, Y is a sulfur atom, Z is a single bond and n is 0 or 1, then all R1 to R5 must not be hydrogen atoms at the same time, and
when X and Y are sulfur atoms, R4 and R5 are hydrogen atom, Z is a single bond and n is 0, then R1 to R3 must not be such that any one of these is a para-positioned chlorine atom while the other two are hydrogen atoms, and their salts and solvates.
In addition, the present invention also provides pharmaceutical compositions comprising any of compounds of the above-mentioned formula (II) and their salts and hydrates, along with pharmaceutically-acceptable carriers.
The present invention further provides ACAT inhibitors, intracellular cholesterol transference inhibitors, blood cholesterol depressants, and inhibitors for macrophage foam cells, which comprise anilide compounds of a formula (IV): 
wherein R1, R2 and R3 are the same or different, and each represents a hydrogen atom, a lower alkyl group, a lower alkoxy group, a halogen atom, a hydroxyl group, a phosphoric acid group, a sulfonamido group, or an optionally-substituted amino group; or any two of R1, R2 and R3 together form an alkylenedioxy group;
R4 and R5 are the same or different, and each represents a hydrogen atom, a lower alkyl group, or a lower alkoxy group: and R4 and R5 may together form a lower alkylene group of which one or more methylene moieties may optionally be substituted by oxygen and/or sulfur atoms;
X represents xe2x80x94NHxe2x80x94, or an oxygen or sulfur atom;
Y represents xe2x80x94NHxe2x80x94, an oxygen or sulfur atom, or a sulfoxide or sulfone group;
Z represents a single bond, or xe2x80x94NR6xe2x80x94;
R6 represents a hydrogen atom or a lower alkylene group; and
n represents an integer of from 0 to 15;
and their pharmaceutically-acceptable salts and solvates, optionally along with pharmaceutically-acceptable carriers. Specifically, the present invention provides medicines for treating, preventing and curing disorders of, for example, hyperlipemia, arteriosclerosis, cervical and cerebral arteriosclerosis, cerebrovascular disorders, ischemic cardiopathy, coronary arteriosclerosis, nephrosclerosis, arteriosclerotic nephrosclerosis, arteriolosclerotic nephrosclerosis, malignant nephrosclerosis, ischemic enterophathy, acute mesenteric vaso-obstruction, chronic intestinal angina, ischemic colitis, aortic aneurysm, and arteriosclerosis obliterans (ASO), which medicines comprise any of compounds of the above-mentioned formula (IV) and their pharmaceutically-acceptable salts and hydrates, optionally along with pharmaceutically-acceptable carriers.
The lower alkyl group as referred to herein is a linear or branched one having from 1 to 8 carbon atoms, preferably from 1 to 6 carbon atoms, and includes, for example, a methyl group, an ethyl group, a n-propyl group, an iso-propyl group, an n-butyl group, an iso-butyl group, a tert-butyl group, a n-pentyl group, and a n-hexyl group. The alkyl moiety in the lower alkoxy group as referred to herein is preferably an alkyl group such as that mentioned hereinabove. The halogen atom is preferably a fluorine, chlorine, bromine or iodine atom. The amino group may optionally be substituted with 1 or 2 substituents. As the substituents for the amino group, preferred are a lower alkyl group such as that mentioned hereinabove; an aryl group such as a phenyl or naphthyl group; and an aralkyl group such as a benzyl or phenethyl group. The aromatic ring in these substituents may further be substituted with any of lower alkyl groups and lower alkoxy groups such as those mentioned hereinabove.
The alkylenedioxy group as referred to herein comprises a linear or branched alkylene group having from 1 to 8 carbon atoms, preferably from 1 to 5 carbon atoms, and has two oxygen atoms as inserted into any desired sites including the both terminals of said alkylene moiety. For example, the group includes a methylenedioxy group and an ethylenedioxy group.
The lower alkyl group for R5 and R6 is preferably one to be selected from those mentioned hereinabove. The lower alkylene group to be formed by R4 and R5 is a linear or branched one having from 1 to 8 carbon atoms, preferably from 2 to 5 carbon atoms, and includes, for example, a methylene group (this forms a vinyl group together with the adjacent carbon atom), a propylene group, a butylene group, and a pentylene group. Of the alkylene group, one or more methylene moieties (each comprising one carbon atom and two hydrogen atoms) may be substituted with oxygen and/or sulfur atoms.
Acid addition salts of compounds (I) of the invention include, for example, inorganic acid salts thereof, such as hydrochlorides, sulfates, nitrates and phosphates; and organic acid salts thereof, such as methanesulfonates, maleates, fumarates, and citrates.
The solvates include, for example, those with solvents used in the production or purification of compounds (I), such as water or alcohols, and the solvents are not specifically defined provided that they do not have any negative influence on the ACAT inhibiting activities of the resulting solvates. As the solvates, preferred are hydrates.
Modes of Carrying out the Invention
Compounds of formulae (I), (II), (III) and (IV) can be produced by any known methods, and the production of said compounds shall not be specifically defined For example, they can be produced according to the following methods.
1. Method of Producing Compounds having a Single Bond as Z
Compounds of formulae (I), (II), (III) and (IV) having a single bond as Z can be produced by reacting a carboxylic acid or its reactive derivative of a general formula (V) with an aniline derivative of a general formula (VI) and then with a 2-substituted benzazole derivative of a general formula (VIII) in that order.
(1) According to the reaction steps mentioned below, a carboxylic acid or its reactive derivative, such as its halide, of formula (V) is reacted with an aniline derivative of formula (VI) to give an aide derivative of a general formula (VII). The resulting derivative (VII) is reacted with a 2-substituted benzazole compound of formula (VIII) to give the intended compound having a sing, bond as Z. 
wherein R7 represents a removable group; and R8 represents a hydroxyl group, or a residue of a reactive derivative for the carboxyl group.
To the reaction of the compound (V) and the compound (VI), applicable is any ordinary means of peptide synthesis. For example, the two compounds are reacted in the presence of a condensing agent to give the intended compound. As the condensing agent, for example, 1-(3xe2x80x2-dimethylaminopropyl)-3-ethylcarbodiimide (WSC) or 1,3-dicyclohexylcarbodinimde (DCC) can be used singly. If desired, said condensing agent can be combined with a condensation activator, such as 1-hydroxybenzotriazole (HOBt) or N-hydroxysuccinide (HOSu). The solvent for the reaction is not specifically defined. For example, usable are dimethylformamide (DMF), methylene chloride, chloroform, tetrahydrofuran and toluene, either singly or as combined. The reaction mode varies, depending on the starting compounds used. In general, however, the starting compounds are reacted with cooling with ice or at higher temperatures but up to the boiling point of the solvent used, for example, at from 0 to 100xc2x0 C., preferably at about room temperature, for from 1 to 30 hours, preferably for from 10 to 20 hours, to finish the reaction. If a highly-reactive carboxylic acid halide is used as the starting compound (V), for example, it may be reacted with the compound (VI) in the presence of a base, such as triethylamine, 4-methylaminopyridine or 14-methylmorpholine, in any ordinary manner. The starting compounds (V) and (VI) are known compounds, For example, the compounds (V) can be obtained through oxidation of a haloalkyl alcohol with a Jones"" reagent or the like into the corresponding carboxylic acid. The compounds (VI) can be obtained through catalytic reduction of a nitrobenzene derivative into the corresponding aniline derivative.
The reaction of the) compound (VII) as obtained in the above-mentioned step and the compound (VIII) may be effected in a solvent in the presence or absence of a base. As the solvent, usable is any of those mentioned above. The base usable herein includes inorganic bases, for example, alkali metal hydroxides such as sodium hydroxide and potassium hydroxide; alkali metal carbonates such as sodium carbonate and potassium carbonate; and alkali metal hydrogencarbonates such as sodium hydrogencarbonate and potassium hydrogencarbonate; as well as organic bases such as pyridine, triethylamine, N,N-diisopropylethylamines, N-methylmorpholine and N,N-dimethylaniline.
(2) Alternatively, according to the reaction steps mentioned below, a 2-substituted benzazole compound of formula (VIII) is reacted with a free carboxylic acid of formula (V) or its derivative as inactivated at its carboxyl moiety to give a benzazole-substituted carboxylic acid derivative of formula (IX). The resulting compound or its reactive derivative, such as its acid halide, of formula (IX) is reacted with an aniline derivative of formula (VI) to give the intended compound having a single bond as Z. 
wherein R7 represents a removable group; and R9 represents a hydroxyl group, or a residue of a derivative for the carboxyl group.
The reaction of the compound (VIII) and the compound (Vxe2x80x2) can be effected in accordance with the second step of the above-mentioned method (1). For this, it is especially desirable to use potassium hydroxide as the base and ethanol as the solvent. The next reaction of the compound (IX) and the compound (VI) can be effected in accordance with the first step of the above-mentioned method (1).
2. Method of Producing Compounds having xe2x80x94NR6xe2x80x94 as Z
(1) Method of Producing Compounds having Hydrogen Atom as R6 
Compounds (I), (II), (III) and (IV) where R4 is hydrogen atom, that is, Z is xe2x80x94NHxe2x80x94, may be produced according to various methods. One example comprises the following reaction steps. 
wherein R7 represents a removing group.
An isocyanate derivative of formula (X) is reacted with an aniline derivative of formula (VI) to give an urea derivative of formula (XI) The resulting urea derivative (XI) is reacted with a 2-substituted benzazole derivative of formula (VII) to give the intended compound having xe2x80x94NR6xe2x80x94 as Z.
In the first step, the compound (X) is reacted with from 1 to 2 equivalents, relative to the compound (X), of the compound (VI) in a solvent to give the compound (XI). The solvent is not specifically defined, but is preferably any of methylene chloride, chloroform, hexane, ether, tetrahydrofuran, toluene, xylene or dimethylformamide. The reaction is effected at from 0xc2x0 C. to the boiling point of the solvent used, for from 1 to 24 hours. The isocyanate derivatives (X) are known compounds, and can be produced, for example, according to a process comprising reacting a carboxylic acid of formula (V) with diphenylphosphoryl azide in the presence of a base (Shioiri et al""s process); or according to a process comprising reacting an acid halide of formula (V) with sodium azide to give an acid azide. The reaction of the compound (XI) and the compound (VIII) may be effected in accordance with the second step of the above-mentioned method 1-(1).
(2) Method of Producing Compounds having Lower Alkyl Group as R6 
Compounds (I), (II), (III) and (IV) where R6 is lower alkyl group may be produced according to various methods. One example comprises the following reaction steps. 
wherein R7 represents a removing group.
An isocyanate derivative of formula (XIII) is reacted with an secondary amine shown as formula (XII) to give an urea derivative of formula (XIV). The resulting urea derivative is reacted with a 2-substituted benzazole derivative of formula (VIII) to give the intended compound having lower alkyl group as R6 in Z.
The reaction between the compound (XII) and the compounds (XII) in the first step may be performed accordance with the reaction between isocyanate derivatives (X) and aniline derivatives (VI) mentioned above, and the reaction between the compounds (XIV) and the compounds (VIII) may be performed accordance with the reaction between amide derivatives (VII) and benzazole compounds (VIII) mentioned above.
Secondary amines shown as the formula (XII) are known compounds, and they may be produced by that amide which is obtained from condensing firstly aminoalcohol and alkylcarbonic acid is reduced.
Also, isocyanate derivatives of the compounds (XIII) may be produced by known methods, for example, the method that corresponded aniline derivatives are made to isocyanate with the one known method using hosgen and so on.
The intermediates and the final compounds obtained in the reaction steps in the above-mentioned methods can be isolated and purified through ordinary purification in organic synthetic chemistry, which includes, for example, filtration, extraction, washing, drying, concentration, recrystallization, and various chromatographic means. For the intermediates, they can be used in the next reaction step without being specifically purified.
In the methods illustrated hereinabove, used are aniline derivatives of formula (VI) as the amines to form the amine moiety in the amido group of the compound of the present invention. However, it is obvious to those skilled in the art that, when arylamine derivatives are used in place of said aniline derivatives, compounds of formulae (I) and (III) of the present invention can be produced.
The compounds of formulae (I), (II), (III) and (IV) thus obtained can be converted into their acid addition salts in any ordinary manner.
If desired, they can be obtained as their solvates, especially their hydrates, with solvents such as those used for reaction or recrystallization.
Specific examples of the compounds of formulae (I), (II), (III) and (IV) obtainable according to the methods mentioned hereinabove are in the following Table 1 to Table 4.
Compounds of formulae (III) and (IV) of the present invention have ACAT inhibiting activities and/or intracellular cholesterol transference inhibiting activities, and are useful as medicines for curing and treating hyperlipemia and those for curing and treating arteriosclerosis in the field of medicine. In particular, since the compounds of the present invention act to selectively inhibit a sub-type of ACAT enzyme existing in blood vessel walls, they are expected to have less side effects than non-selective ACAT inhibitors. Thus, the compounds of the invention are favorable as active ingredients in medicines.
The pharmaceutical composition of the present invention comprises, as the active ingredient, any of compounds of formulae (III) and (IV) and their acid addition salts and solvates, either singly or along with any other pharmaceutically-acceptable excipients, binders, carriers or diluents, and can be formulated into various preparations such as tablets, capsules, granules,, powders, injections and suppositories. To formulate these preparations, employable are any known methods. For example, to formulate oral preparations, the active ingredient of compounds (III) and (IV) is combined with an excipient such as starch, mannitol or lactose; a binder such as sodium carboxymethyl cellulose or hydroxypropyl cellulose; a disintegrator such as crystalline cellulose or calcium carboxymethyl cellulose; a lubricant such as talc or magnesium stearate; and a flowability improver such as light silicic anhydride.
The pharmaceutical composition of the present invention is administered to patients either orally or parenterally.
The dose of the pharmaceutical composition of the invention shall vary, depending on the body weight, the age, the sex and the condition of the patient. Preferably, however, the dose is generally from 1 to 1000 mg/adult/day, more preferably from 5 to 200 mg/adult/day, in terms of the active ingredient of compounds (III) and (IV) existing in the composition, and the composition of said dose is administered once to three times a day.
The following Test Example is to demonstrate the ACAT inhibiting activities of compounds of formulae (III) and (IV) of the invention.
Rabbits were fed with 1% cholesterol feed for 8 weeks. Microsomes were prepared from the thoracoaorta of the rabbits in an ordinary manner, and suspended in 0.15 M phosphate buffer (pH 7.4) to prepare blood vessel wall-derived enzyme liquids, Small intestine-derived enzyme liquids were prepared from the small intestines of rabbits fed with ordinary feed. To determine the ACAT inhibiting activities of test compounds, employed herein was a modified method of J. G. Heider (see J. Lipid Res., 24, 1127-1134, 1983) Briefly, 2 xcexcl of a test compound as dissolved in dimethylsulfoxide (DMSO) was added to 88 xcexcl of 0.15 M phosphate buffer (pH 7.4) containing 14C-oleoyl-CoA (40 xcexcM, 60000 dpm) and bovine serum albumin (2.4 mg/ml), and incubated therein at 37xc2x0 C. for 5 minutes. 10 xcexcl of the enzyme liquid was added to the resulting solution and reacted at 37xc2x0 C. for 5 minutes (for small intestine-derived enzyme liquids, for 3 minutes), and thereafter 3 ml of chloroform/methanol (2/1) and 0.5 ml of 0.04 N HCl were added thereto to stop the reaction. After the reaction, lipids were extracted from the reaction mixtures. The solvent layer was concentrated to dryness, then dissolved in hexane, spotted on a TLC plate (produced by Merck), and developed thereon with a developer of hexane/ether/acetic acid (75/25/1) The radioactivity of the thus-fractionated cholesterol ester fraction was measured with BAS2000 (produced by Fuji Film). Comparing the data with those of the control to which had been added only DMSO, IC50 was obtained. The results are shown in Table 5.
As comparative compounds, 5-[2-(2-(4-fluorophenyl)ethyl)-3-(1-methyl-1H-imidazol-2-yl) -2H-benzopyran-6-yl]oxy-2,2-dimethyl-N-(2,6-diisopropylphenyl)pentanamide described in WO92/09582 (control (1)), N-(2, 6-diisopropylphenyl) -2-(tetradecylthio)acetamide described in WO92/09572 (control (2)), N-butyl-N-[2-(3-(5-ethyl-4-phenyl-1H-imidazol-1-yl)propoxy)-6-methylphenyl]urea described in EP-A 477,778 (control (3)), and N4-[5-(4,5-diphenyl-1H-imidazol-2-ylthio)pentyl]-N-heptyl-2-benzoxazolamine described in WO93/23392 (control (4)) were tested in the same manner as above, and the data obtained are also shown in Table 5.
Test Example 2
Deterioration Action of Cholesterol
A deterioration action of cholesterol in plasma was tested with BIO F1B hybrid syrian hamster (Mark C. K. et al., Atheorosclerosis, 91 (1991) 35-49: abbreviated as FIB hamster in the following) which has an accumulation of Cholesterol in blood vessel by cholesterol load. That is 4 groups of FIB hamster (male, 8 weeks old, about 100 g weight) were made, which had five hamsters respectively, and the hamsters were fed with the below-described feeds corresponding to each group for 4 weeks. At the 4th week from beginning feed, blood of the hamsters were drawn from their jugulars; and values of total cholesterols in plasma were measured with the enzyme method. The results are shown in Table 6.
Test Example 3
ACAT inhibiting activities in J774 cell and HepG2 cell (Anti-foaming activities).
J774 cell and HepG2 cell were sown on 24-holes-plates, and J774 cell was cultured with DMEM culturing solution (including 10% fetal bovine serum) and HepG2 cell was cultured with MEM culturing solution (including 10% fatal bovine serum) and they were incubated with 5% CO2 at 37xc2x0 C. for 24 hours. And then each culturing solutions was changed to 0.5 ml of the culturing solution including 10 xcexcg/ml of 25-OH cholesterol and specimen, and each cell was further cultured for 18 hours. After redwing the medium, each cultured cell was washed with PBS two ties, and then extracted with 1.5 ml of hexane:iso-propanol (3:2), and concentrated and dried. Each extracted substance was dissolved into iso-propanol including 0.2 ml of 10% Triton X-100. And total cholesterol (TC) of each was measured with Cholesterol E Test Wako (Wako Junyaku Co.), and free cholesterol (EC) of each was measured with Free Cholesterol E Test Wako (Wako Junyaku Co.). The residue of extracting was dissolved with 0.25 ml of 2N NaOH at 37xc2x0 C. for 30 minutes, and an amount of protein was measured with BCA Protein Assay Reagent (Pierce). The amount of cholesterol ester per protein was calculated based on a difference between TC and FC, and IC50 was calculated in contrast with control. The results are shown in Table 7.
As the above-mentioned results, the compounds of the present invention has the superior inhibition for ACAT, in particular, since the compounds of the present invention act to selectively inhibit a sub-type of the ACAT enzyme existing in blood vessel wall, it is found that they are erected as medicines for curing arteriosclerosis which have less side effects than non-selective CAT inhibitions And also, the compounds of the present invention has an activity to deteriorate cholesterol which depends on dose.